Tetraguaiacol Reaction

Introduction: Many vital chemical reactions do not naturally happen at fast enough rates to maintain life. Enzymes are a type of protein that speed up these reactions because they are catalytic, meaning they increase the rate of the reaction without being consumed by it (Freeman, 2014, p. 54). In 1894, Emil Fischer proposed the “lock-and-key” method in order to explain how enzymes work. As the name suggests, the enzyme represents the lock as the substrate represents the key. A substrate is the reactant the enzyme binds to that the enzyme then quickly transforms into a product (Campbell, 2001, p. 151). The substrate only binds to a particular region of the enzyme called the active site. At the active site, hydrogen bonds or other weaker intermolecular

Hydrogen peroxide is a toxic by-product of many crucial metabolic reactions for aerobic organisms. Lastly, a colorless dye called guaiacol is added to the reaction so that it can bind to peroxidase, get oxidized while hydrogen peroxide is reduced to water, and form a brown tetraguaiacol. That is an example of an oxidation-reduction reaction, which can be monitored using a spectrophotometer due to the brown nature of tetraguaiacol. With a LabQuest attached to a spectrophotometer, absorbance of the enzyme/substrate solution is monitored versus time when exposed to different conditions.
In the first experiment, temperature affects the rate enzyme activity, and the more temperature increases, the faster the reaction will proceed until the optimum temperature is reached. After that, enzyme activity will decrease. Furthermore, hydroxylamine and hydrogen peroxide have extremely similar structures. In the second experiment, hydroxylamine affects peroxidase’s activity. The more hydroxylamine, the less enzyme activity will occur due to the competitive inhibition of the

In part B of the experiment, the amount of peroxidase to use for the rest of the experiment was standardized, and it was concluded that 250μl of peroxidase should be used in the experiment for optimal results. In part C of the experiment, the effect of temperature was tested. Six cuvettes were numbered, and three mL of peroxidase was added to a test tube to then be placed in a hot/boiling water bath for 15 minutes. After the test tube was taken out and let cooled, 250μl of the boiled water was added to cuvette six. Cuvette two was placed in ice, cuvette four was placed in the 32°C water bath, and cuvette five was placed in a 48°C water bath for ten minutes. After the ten minutes was up, 1000μl of buffer was added to cuvettes one through six, 500 mL of guaiacol was added to all cuvettes as well, and 250mL of normal peroxidase was added to cuvettes two through five. Cuvette one was used to calibrate the spectrophotometer. All of these values can be seen in the table